CN105519439A - Ginseng rapid proliferation culture medium and preparation method thereof - Google Patents
Ginseng rapid proliferation culture medium and preparation method thereof Download PDFInfo
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- CN105519439A CN105519439A CN201610011344.1A CN201610011344A CN105519439A CN 105519439 A CN105519439 A CN 105519439A CN 201610011344 A CN201610011344 A CN 201610011344A CN 105519439 A CN105519439 A CN 105519439A
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- ginseng
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- fast breeding
- caseinhydrolysate
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Engineering & Computer Science (AREA)
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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a ginseng rapid proliferation culture medium, and relates to the technical field of ginseng tissue culture. According to the invention, MS culture medium is adopted as a base culture medium, and 6-BA (6-benzylamino adenine), 2,4-D (2,4-dichlorophenoxyacetic acid), CH (casein hydrolyzate), agar powder and white granulated sugar are added. The ginseng rapid proliferation culture medium provided by the invention has the advantages of short proliferation time and large proliferation coefficient.
Description
Technical field
The invention belongs to ginseng tissue culture technique field, particularly a kind of ginseng fast breeding medium, also relate to a kind of compound method of ginseng fast breeding medium.
Background technology
Ginseng is Angiospermae Dicotyledoneae Archichlamydeae Araliaceae, herbaceos perennial, be grown on thick forest, be distributed in Heilungkiang, Jilin, Liaoning and northern Hebei remote mountains more, containing more than 10 kinds of panaxosides, and the fast alcohol of ginseng, several amino acids and vitamin, be famous and precious Chinese medicinal herbs.
But the at present ginseng tissue indefinite bud poor growth of cultivating, generation time is long, and growth coefficient is low, is difficult to meet the requirement of market, scientific research and species conservation.
Summary of the invention
There are the problems referred to above based on prior art, the invention provides a kind of ginseng fast breeding medium, it is medium based on MS medium, and be added with 6-BA(6-benzyl aminoadenine), 2,4-D(2,4-dichlorphenoxyacetic acid), CH(caseinhydrolysate), agar powder and white granulated sugar, it is short that ginseng fast breeding medium provided by the invention has generation time, the advantage that growth coefficient is large.
A kind of ginseng fast breeding medium, medium based on MS medium, and be added with 6-BA(6-benzyl aminoadenine), 2,4-D(2,4-dichlorphenoxyacetic acid), CH(caseinhydrolysate), agar powder and white granulated sugar, caseinhydrolysate contains several amino acids, can provide ginseng indefinite bud growth needs, partial amino-acid also plays the effect regulating cell separation differentiation, promotes adventitious bud proliferation.
The concentration of 6-BA (6-benzyl aminoadenine) is 0.5-1.5mg/L, 2,4-D(2,4-dichlorphenoxyacetic acid) concentration be 0.5-2mg/L, CH(caseinhydrolysate) concentration be 200-500mg/L, the concentration of agar powder is 8 ~ 11g/L, and the concentration of sucrose is 25 ~ 35g/L.
The concentration of 6-BA (6-benzyl aminoadenine) is 0.8-1.2mg/L, 2,4-D(2,4-dichlorphenoxyacetic acid) concentration be 1-1.5mg/L, CH(caseinhydrolysate) concentration be 350-450mg/L, the concentration of agar powder is 9.5 ~ 10.5g/L, and the concentration of sucrose is 29 ~ 31g/L.
The pH value of described ginseng fast breeding medium is 6.3 ~ 6.9.
The pH value of described ginseng fast breeding medium is 6.7, is applicable to ginseng adventitious bud proliferation close to neutral culture environment.
Described caseinhydrolysate is the acid hydrolyzed casein using hydrochloric acid hydrolysis.
A compound method for ginseng fast breeding medium, it comprises the following steps:
Step S10 dissolves preparation medium, dissolves MS basal medium each composition, and add 6-BA(6-benzyl aminoadenine with distilled water), 2,4-D(2,4-dichlorphenoxyacetic acids), CH(caseinhydrolysate), agar powder and white granulated sugar;
Step S20 high-temperature sterilization, the medium prepared by step S10 is put in high-pressure sterilizing pot, in 120 ~ 121 DEG C of high-temperature sterilizations 20 ~ 21 minutes, cooled and solidified.
Described step S10 also comprises step S11 adjust ph, uses HCL and KOH titration to regulate Medium's PH Value to 6.7.
Embodiment
Embodiment one: preparation ginseng fast breeding medium.
Ginseng fast breeding culture medium prescription according to the MS culture medium prescription shown in table 1 and table 2, prepares ginseng fast breeding medium of the present invention.
Weigh in the balance and get each composition in MS medium and 6-BA(6-benzyl aminoadenine), 2,4-D(2,4-dichlorphenoxyacetic acid), CH(caseinhydrolysate), agar powder and white granulated sugar, be placed in triangular flask, add appropriate distilled water, stirring and dissolving, and adjust ph to 6.7, be settled to 1000mL.Triangular flask is sealed, is placed in high-pressure sterilizing pot, in 121 DEG C of sterilizing 20min, is then placed in superclean bench and naturally cools, for subsequent use.
Being divided by medium when will be cooled to 50-60 DEG C is filled in tissue culture flasks, dries in the air cool, by tissue cultures bottle closure after it solidifies, for subsequent use.
The composition of table 1MS medium and consumption thereof.
The composition of table 2 ginseng fast breeding medium and consumption thereof.
Embodiment two: the test of ginseng fast breeding medium of the present invention
Ginseng fast breeding culture medium prescription according to the MS culture medium prescription shown in table 1 and table 3, with reference to the medium preparation method described in embodiment one, prepares the ginseng fast breeding medium of each experimental group, control group respectively.
The composition of table 3 ginseng fast breeding medium and consumption thereof.
Choose in superclean bench growth normal, without the ginseng indefinite bud of brownization, indefinite bud is taken out, with aseptic operation cutter, indefinite bud cutting is slit into single indefinite bud, being inoculated in the medium of each experimental group, control group shown in table 3, is then that 14h illumination/10h is dark, intensity of illumination is 2300lux, temperature is cultivated under being the condition of 23 ± 2 DEG C in light dark period.Statistics propagation ginseng value coefficient and record grow the time of new indefinite bud the soonest afterwards to cultivate 35 days.
Table 4 fleece-flower root Shoot propagation medium test result.
Note: the 1. rear total bud number/inoculation bud number of growth coefficient=increment in table; On average breed each mean value repeating the time of breeding a sprouting the soonest that the used time refers to control group or experimental group the soonest.
From the result of the test of table 4, adopt ginseng fast breeding medium provided by the invention growth coefficient can be brought up to more than 5, simultaneously can find out that the adventitious bud proliferation of caseinhydrolysate to ginseng does not have inducing action from the contrast of control group 3 and experimental group, but but can significantly promote that ginseng is not defined in propagation, improve the effect of plant hormone widely, from the mutual contrast of experimental group, it can also be seen that plant hormone interpolation is excessive in addition can affect facilitation effect.
Claims (8)
1. a ginseng fast breeding medium, is characterized in that, medium based on MS medium, and is added with 6-BA(6-benzyl aminoadenine), 2,4-D(2,4-dichlorphenoxyacetic acids), CH(caseinhydrolysate), agar powder and white granulated sugar.
2. a kind of ginseng fast breeding medium according to claim 1, it is characterized in that, the concentration of 6-BA (6-benzyl aminoadenine) is 0.5-1.5mg/L, 2,4-D(2,4-dichlorphenoxyacetic acid) concentration be 0.5-2mg/L, CH(caseinhydrolysate) concentration be 200-500mg/L, the concentration of agar powder is 8 ~ 11g/L, and the concentration of sucrose is 25 ~ 35g/L.
3. a kind of ginseng fast breeding medium according to claim 2, it is characterized in that, the concentration of 6-BA (6-benzyl aminoadenine) is 0.8-1.2mg/L, 2,4-D(2,4-dichlorphenoxyacetic acid) concentration be 1-1.5mg/L, CH(caseinhydrolysate) concentration be 350-450mg/L, the concentration of agar powder is 9.5 ~ 10.5g/L, and the concentration of sucrose is 29 ~ 31g/L.
4. according to claims 1 to 3 one of them described in a kind of ginseng fast breeding medium, it is characterized in that, the pH value of described ginseng fast breeding medium is 6.3 ~ 6.9.
5. described a kind of ginseng fast breeding medium according to claim 4, it is characterized in that, the pH value of described ginseng fast breeding medium is 6.7.
6. according to claims 1 to 3 one of them described in a kind of ginseng fast breeding medium, it is characterized in that, described caseinhydrolysate uses the acid hydrolyzed casein of hydrochloric acid hydrolysis.
7. a compound method for ginseng fast breeding medium, is characterized in that, it comprises the following steps:
Step S10 dissolves preparation medium, dissolves MS basal medium each composition, and add 6-BA(6-benzyl aminoadenine with distilled water), 2,4-D(2,4-dichlorphenoxyacetic acids), CH(caseinhydrolysate), agar powder and white granulated sugar;
Step S20 high-temperature sterilization, the medium prepared by step S10 is put in high-pressure sterilizing pot, in 120 ~ 121 DEG C of high-temperature sterilizations 20 ~ 21 minutes, cooled and solidified.
8. the compound method of a kind of ginseng fast breeding medium according to claim 7, it is characterized in that, described step S10 also comprises step S11 adjust ph, uses HCL and KOH titration to regulate Medium's PH Value to 6.7.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610011344.1A CN105519439A (en) | 2016-01-09 | 2016-01-09 | Ginseng rapid proliferation culture medium and preparation method thereof |
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| CN201610011344.1A CN105519439A (en) | 2016-01-09 | 2016-01-09 | Ginseng rapid proliferation culture medium and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069770A (en) * | 2016-07-08 | 2016-11-09 | 徐伟明 | A kind of promote the tissue culture medium (TCM) of saponin content in Radix Ginseng |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657095A (en) * | 2012-05-25 | 2012-09-12 | 东北师范大学 | Efficient tissue culture and regeneration method for ginseng |
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2016
- 2016-01-09 CN CN201610011344.1A patent/CN105519439A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657095A (en) * | 2012-05-25 | 2012-09-12 | 东北师范大学 | Efficient tissue culture and regeneration method for ginseng |
Non-Patent Citations (3)
| Title |
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| 唐巍等: "人参悬浮细胞系的建立及其生长特性的研究", 《生物技术》 * |
| 杨震: "盐酸快速水解酪蛋白工艺研究", 《食品研究与开发》 * |
| 王逸群等: "西洋参组织培养及愈伤组织中人参皂苷Rg1和Rb1含量分析", 《漳州师范学院学报（自然科学版）》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069770A (en) * | 2016-07-08 | 2016-11-09 | 徐伟明 | A kind of promote the tissue culture medium (TCM) of saponin content in Radix Ginseng |
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